Inhibition of phosphatidylinositol 4-phosphate (PI4P) and phosphatidylinositol 4,5-bisphosphate (PI4,5P(2)) synthesis by phenylarsine oxide (PAO) inhibits both [3H]-noradrenaline ([3H]-NA) and [14C]-glutamate ([14C]-glu) exocytosis from streptolysin-O (SLO)-perforated synaptosomes. When PI4,5P(2) is blocked by an antibody or chelated by neomycin, neurotransmitter exocytosis again is inhibited. Also, when phosphoinositide (PI) synthesis is indirectly decreased by shunting phosphatidic acid (PA) synthesis into phosphatidylbutanol production, both [14C]-glutamate and [3H]-noradrenaline exocytosis are inhibited. All of these results indicate that PI4,5P(2) is necessary for exocytosis of both synaptic vesicles (SVs) and dense core vesicles (DCVs).